FIG. 1.

(A) Genetic map of AAV. Transcripts and protein-encoding regions for the three AAV promoters are shown. The promoters, intron donor (D) and acceptors (A1 and A2), and ITRs are mapped with their nucleotide designations. (B) The ratios of spliced to unspliced RNAs generated from the different AAV promoters are different during both wild-type AAV infection and transfection of full-length AAV plasmid constructs. 293 cells were either infected with AAV2 (MOI, 10) or transfected with either psub201 or ΔITR in the absence (−) or presence (+) of adenovirus (AD) (MOI, 10). A map of these constructs (not to scale) is shown. Ten micrograms of total RNA from infected or transfected cells was protected by the RP probe, which is depicted in relation to the AAV genome. A representative experiment is shown, and the AAV-specific RNA bands are indicated on the right. Multiple bands protected by P40 unspliced and spliced RNA likely reflect the use of multiple initiation sites within these regions (27). Quantitations of the ratio of spliced to unspliced RNAs specifically derived from the P5+P19 and P40 promoters are shown. All of the values are averages of the results of at least three separate experiments, with standard deviations in parentheses. Previous work has shown (27) that the ratios of spliced to unspliced AAV RNA increase through the course of infection; lane 2 shows the maximal levels of spliced products seen at late times during infection. N/D, not determined